Protein on polymers

The bell-shape-profile concept for protein adsorption may be a useful guideline for researchers when they consider the adsorptive behavior of proteins on polymers with different degrees of hydrophilicity. As will be discussed in later Sect. (2.3), when material surfaces carry ionic groups, the contribution from... 

Gel Permeation Chromatography. Tc-labeled proteins on polymer-based column packing can be purified from precursor reagents and unbound radiolabeling. The first eluted macromolecules are separated according to their size from the smaller molecules. 

The combination of XPS with another technique (AFM and, or radiolabeling) to characterize adsorbed phases is further illustrated for adsorption of proteins on polymers and on self-assembled monolayers,and for mixed Langmuir-Blodgett films of DPPC and surfactin deposited on mica. ... 

Morin C, Hitchcock AP, Cornelius RM, Brash JL, Urquhart SG, A. Scholl A, Doran A. (2004) Selective adsorption of protein on polymer surfaces studied hy soft X-ray photoemission electron microscopy. ]El Spec RelPhenom 137—140 785—794. 

Adsorption and diffusion phenomena at or within polymer systems were investigated by surface sensitive ATR-FTIR spectroscopy. For a systematic description, a study was made of (1) the competitive adsorption and desorption behaviour of proteins on polymer surfaces, (2) swelling of hydrophilic polymers by water molecules, which can be accompanied by conformational changes, and (3) induced orientational changes of hydrophobically modified polypeptides by apolar solvents. 10 refs. 

Size exclusion was first noted in the late fifties when separations of proteins on columns packed with swollen maize starch were observed (Lindqvist and Storgards, 1955 Lathe and Ruthven, 1956). The run time was typically 48 hr. With the advent of a commercial material for size separation of molecules, a gel of cross-linked dextran, researchers were given a purposely made material for size exclusion, or gel filtration, of solutes as described in the classical work by Porath and Flodin (1959). The material, named Sephadex, was made available commercially by Pharmacia in 1959. This promoted a rapid development of the technique and it was soon applied to the separation of proteins and aqueous polymers. The work by Porath and Flodin promoted Moore (1964) to apply the technique to size separation, gel permeation chromatography of organic molecules on gels of lightly cross-linked polystyrene (i.e., Styragel). 

Many proteins and polymers have been analyzed on SynChropak GPC and CATSEC columns. Table 10.6 lists some of the published applications. The use of a surfactant to analyze the caseins in milk is illustrated in Eig. 10.12. Viruses have also been analyzed on SynChropak GPC columns, as seen in the chromatogram from Dr. Jerson Silva of the University of Illinois (Pig. 10.13). Dr. Nagy and Mr. Terwilliger analyzed cationic polymers on a series of CATSEC columns using differential viscometry as detection (Pig. 10.14) (9). 

Researchers are facing difficulties in improving the properties and response rates of chemomechanical andelectrochemomechanical systems based on polymer gels or proteins that are intended to be used as actuators in robotics. Lack of mechanical toughness and long-term durability are other problems to be solved. A basic improvement in the low efficiency... 

In this chapter we focus on polymers, from the plastics that are essential in our everyday lives to the proteins and DNA that are the materials of life. To understand the chemishy of polymers, we need to understand how monomers are able to link together into long chains. We begin with a description of the various monomers that are the most important starting materials for industrial and natural polymers. 

Disperse systems can also be classified on the basis of their aggregation behavior as molecular or micellar (association) systems. Molecular dispersions are composed of single macromolecules distributed uniformly within the medium, e.g., protein and polymer solutions. In micellar systems, the units of the dispersed phase consist of several molecules, which arrange themselves to form aggregates, such as surfactant micelles in aqueous solutions. 

Other reviews of multidimensional separations have been published. These include a book on polymer characterization by hyphenated and multidimensional techniques (Provder et al., 1995), a review on polymer analysis by 2DLC (van der Horst and Schoenmakers, 2003), and two reviews on two-dimensional techniques in peptide and protein separations (Issaq et al., 2005 Stroink et al., 2005). Reviews on multidimensional separations in biomedical and pharmaceutical analysis (Dixon et al. 2006) and multidimensional column selectivity (Jandera, 2006) were recently published. Suggested nomenclature and conventions for comprehensive multidimensional chromatography were published in 2003 (Schoenmakers et al., 2003), and a book chapter in the Advances in Chromatography series on MDLC was published in 2006 (Shalliker and Gray 2006). 

The application of polymer monoliths in 2D separations, however, is very attractive in that polymer-based packing materials can provide a high performance, chemically stable stationary phase, and better recovery of biological molecules, namely proteins and peptides, even in comparison with C18 phases on silica particles with wide mesopores (Tanaka et al., 1990). Microchip fabrication for 2D HPLC has been disclosed in a recent patent, based on polymer monoliths (Corso et al., 2003). This separation system consists of stacked separation blocks, namely, the first block for ion exchange (strong cation exchange) and the second block for reversed-phase separation. This layered separation chip device also contains an electrospray interface microfabricated on chip (a polymer monolith/... 

Akagi T, Kaneko T, Kida T et al (2006) Multifunctional conjugation of proteins on/into coreshell type nanoparticles prepared by amphiphilic poly(y-glutamic acid). J Biomater Sci Polym Ed 17 875-892... 

Tamada Y, Ikada Y (1993) Effect of preadsorbed proteins on cell adhesion to polymer surfaces. J Colloid Interface Sci 155 334-339... 

Reverse-phase HPLC (RP-HPLC) separates proteins on the basis of differences in their surface hydophobicity. The stationary phase in the HPLC column normally consists of silica or a polymeric support to which hydrophobic arms (usually alkyl chains, such as butyl, octyl or octadecyl groups) have been attached. Reverse-phase systems have proven themselves to be a particularly powerful analytical technique, capable of separating very similar molecules displaying only minor differences in hydrophobicity. In some instances a single amino acid substitution or the removal of a single amino acid from the end of a polypeptide chain can be detected by RP-HPLC. In most instances, modifications such as deamidation will also cause peak shifts. Such systems, therefore, may be used to detect impurities, be they related or unrelated to the protein product. RP-HPLC finds extensive application in, for example, the analysis of insulin preparations. Modified forms, or insulin polymers, are easily distinguishable from native insulin on reverse-phase columns. 

LOV MOLECULAR WEIGHT MODEL COMPOUNDS. The mechanisms of radiation effects on polymers are frequently investigated by studies of low molecular weight model compounds. Analysis of the chemical reactions is much easier than with high molecular weight polymers. Thus, N-acetyl amino acids can be studied as model compounds for poly(amino acid)s and hence for proteins. 

Figure 3.15 Chromatogram of fibre-type proteins on polystyrene gels having different pore sizes. Column A, PLRP-S 300 A, 15 cm x 4.6 mm i.d. B, PLRP-S 1000 A (polystyrene gel), 15 cm x 4.6 mm i.d. eluent, 15 min linear gradient from 20% of 0.25% trifluoroacetic acid to 60% of 0.25% trifluoro-acetic acid in 95% aqueous acetonitrile flow rate, 1.0 ml min-1 detection, UV220 nm. Peaks 1, collagen (Mr 120 000) and 2, fibrinogen (Mr 340 000). (Reproduced by permission from Polymer Laboratories data)...

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